title: An improved simplified high-sensitivity quantification method for determining brassinosteroids in different tissues of rice and Arabidopsis.
abstract: Quantification of brassinosteroids is essential and extremely important to study the molecular mechanisms of their physiological roles in plant growth and development. Herein, we present a simple, material and cost-saving high-performance method for determining endogenous brassinosteroids (BRs) in model plants. This new method enables simultaneous enrichment of a wide range of bioactive BRs such as brassinolide, castasterone, teasterone, and typhasterol with ion exchange solid-phase extraction and high-sensitivity quantitation of these BRs based on isotope dilution combined with internal standard approach. For routine analysis, the consumption of plant materials was reduced to one-twentieth of previously reported and the overall process could be completed within 1 day compared with previous 3 to 4 days. The strategy was validated by profiling BRs in different ecotypes and mutants of rice (Oryza sativa) and Arabidopsis (Arabidopsis thaliana), and the BR distributions in different model plants tissues were determined with the new method. The method allows plant physiologists to monitor the dynamics and distributions of BRs with 1 gram fresh weight of model plant tissues, which will speed up the process for the molecular mechanism research of BRs with these model plants in future work.
authors.affiliation: National Centre for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Peiyong
authors.initials: P
authors.lastname: Xin
authors.forename: Jijun
authors.initials: J
authors.lastname: Yan
authors.forename: Jinshi
authors.initials: J
authors.lastname: Fan
authors.forename: Jinfang
authors.initials: J
authors.lastname: Chu
authors.forename: Cunyu
authors.initials: C
authors.lastname: Yan
mesh_terms: D017360:Arabidopsis / Q000737:chemistry* / Q000235:genetics; D060406:Brassinosteroids / Q000032:analysis* / Q000302:isolation & purification; D002777:Cholestanols / Q000032:analysis; D002852:Chromatography, Ion Exchange / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D009154:Mutation; D012275:Oryza / Q000737:chemistry* / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction; D013260:Steroids, Heterocyclic / Q000032:analysis
chemical_list: D060406:Brassinosteroids; D002777:Cholestanols; D013260:Steroids, Heterocyclic; C083545:castasterone; C023623:brassinolide